Papillomaviruses are emerging as possible agents of cancer in man and have been associated with numerous animal cancers. The small amount of information available on their molecular biology suggess functional differences in maintenance of cell transformation from that of the well characterized DNA tumor viruses. This project has been designed to provide insights into the functional relationshbips between the papillomavirus genome and host cell using the three papillomaviruses thus far shown to be active in vitro. The specific aims of the project are to: (i) molecularly clone bovine type 1 and type 2 papillomavirus (BPV-1 and BPV-2) and deer fibromavirus (DFV), also a papillomavirus, NDAs and subgenomic fragments into E. coli K12 using the certified plasmid vector pBR322; (ii) assay biological activity of th putative transforming and nontransforming replicative regions of the BPV-1, BPV-2, and DFV genomes. Transforming regions of the genomes will be assayed on susceptible mouse cells. Nontransforming-replicating fragments will be assayed by blot-transfer hy bridization of DNA obained from thymidine kinas (TK) deficient mouse cells cotransformed with replicactive papillomavirus DNA and herpes virus TK gene. The replication origin of papillomavirus DNAs will be mapped by electron microscopy using replicating nonintegrated viral sequences present in high copy number in transformed cells; (iii) characterize virus-specific transforming proteins by immunoprecipitation of polypeptides from transformed cells using sera from tumor bearing animals. Map virus-specific transcripts on the viral genomes by hy bridization of viral RNA sequences to separated restriction enzyme cleavange fragments of virus DNA and translate viral mRNAs in vitro or in Xenopus oocytes; (iv) vigorously assess the state of papillomavirus sequences in virus-induced tumors, tumor cell lines, and cell transformed in culture by in situ hybridization and blot-transfer analysis; (v) determine the DNA sequence of the origin DNA replication and transforming region and eventually the entire genome of either BPV-1, BPV-2, or DFV USING M13 cloning and didseoxy sequencing techniques.